title: Myosin II regulates activity dependent compensatory endocytosis at central synapses.
abstract: Recent evidence suggests that endocytosis, not exocytosis, can be rate limiting for neurotransmitter release at excitatory CNS synapses during sustained activity and therefore may be a principal determinant of synaptic fatigue. At low stimulation frequencies, the probability of synaptic release is linked to the probability of synaptic retrieval such that evoked release results in proportional retrieval even for release of single synaptic vesicles. The exact mechanism by which the retrieval rates are coupled to release rates, known as compensatory endocytosis, remains unknown. Here we show that inactivation of presynaptic myosin II (MII) decreases the probability of synaptic retrieval. To be able to differentiate between the presynaptic and postsynaptic functions of MII, we developed a live cell substrate patterning technique to create defined neural circuits composed of small numbers of embryonic mouse hippocampal neurons and physically isolated from the surrounding culture. Acute application of blebbistatin to inactivate MII in circuits strongly inhibited evoked release but not spontaneous release. In circuits incorporating both control and MIIB knock-out cells, loss of presynaptic MIIB function correlated with a large decrease in the amplitude of evoked release. Using activity-dependent markers FM1-43 and horseradish peroxidase, we found that MII inactivation greatly slowed vesicular replenishment of the recycling pool but did not impede synaptic release. These results indicate that MII-driven tension or actin dynamics regulate the major pathway for synaptic vesicle retrieval. Changes in retrieval rates determine the size of the recycling pool. The resulting effect on release rates, in turn, brings about changes in synaptic strength.
authors.affiliation: Departments of Anatomy and Neurobiology and Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri 63110, and Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts 02138.
authors.forename: Indra
authors.initials: I
authors.lastname: Chandrasekar
authors.forename: James E
authors.initials: JE
authors.lastname: Huettner
authors.forename: Stephen G
authors.initials: SG
authors.lastname: Turney
authors.forename: Paul C
authors.initials: PC
authors.lastname: Bridgman
mesh_terms: D000818:Animals; D004705:Endocytosis / Q000502:physiology*; D006624:Hippocampus / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D046529:Microscopy, Electron, Transmission; D024462:Myosin Type II / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism; D018377:Neurotransmitter Agents / Q000378:metabolism*; D018408:Patch-Clamp Techniques; D013569:Synapses / Q000378:metabolism*; D009435:Synaptic Transmission / Q000502:physiology*; D013572:Synaptic Vesicles / Q000378:metabolism
chemical_list: D018377:Neurotransmitter Agents; D024462:Myosin Type II